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. 2018 Jan 30;12:17. doi: 10.3389/fncel.2018.00017

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Copyright © 2018 Boido, De Amicis, Valsecchi, Trevisan, Ala, Ruegg, Hettwer and Vercelli.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Analysis of diaphragm NMJs. (A–C) Evaluation of NMJ maturation by BTX-immunostaining: NT-1654 injection partially stimulates NMJ maturation in treated SMA (B) compared to PBS SMA (A), as quantified in (C). Mature endplates are indicated by arrowheads. (D–F) Double immunostaining against BTX (red) and NF (green) is employed for analyzing NF accumulation into the plaques: NT-1654 SMA (E) display a significantly lower number of engulfed NMJs (arrows) compared to PBS SMA (D), as also quantified in (F). Unpaired T-test, ^*P < 0.05. Scale bar = 25 μm in (A,B,D,E).